A new simple method to introduce marker-free deletions in the Bacillus subtilis genome

作者:Morimoto Takuya; Ara Katsutoshi; Ozaki Katsuya; Gasawara Naotake*
来源:Genes & Genetic Systems, 2009, 84(4): 315-318.
DOI:10.1266/ggs.84.315

摘要

A genetic tool to introduce marker-free deletions is essential for multiple manipulations of genomes. We report a simple and efficient method to create marker-free deletion mutants of Bacillus subtilis through transformation with recombinant PCR products, using the Escherichia coli mazF gene encoding an endoribonuclease that cleaves free mRNAs as a counter-selection tool. Our method will be applicable to any bacterium in which introduction of the mazF cassette into the genome by double crossover homologous recombination is possible.

  • 出版日期2009-8